How n pheromone studies omplement txonomy: lik eetle exmple (Coleopter: Elteride) Miklós Tóth 1, József Vuts 1, Lorenzo Furln 2, Till Tolsh 3 Amáli Xvier, Teodor Toshov 5, Mitko Suhev 5, István Szrukán 6, Venymin Ytsynin 7 1 Plnt Protetion Institute, HAS, Budpest, Hungry; 2 Deprtment of Agronomy, Entomology, Pdov University, Agripolis, Itly; 3 Univ. Hohenheim, Inst. Zool. Fg. Tierökologie, Grenstr. 3, D-7593 Stuttgrt Germny; DRAEDM, Porto, Portugl; 5 Zoology Institute, BAS, Sofi, Bulgri; 6 Agriulturl University, Dereen, Hungry; 7 Krsnodrskiy NIISKh im. P.P. Lukynenko, Krsnodr 12, Russi
Wireworms, the lrve of lik (Coleopter: Elteride) re importnt soil-dwelling polyphgous pests ll over the world www.nturmediterrneo.om photoshelter.om
Trditionl forest nd monitoring involves lour-intensive soil smpling methods,
nd to otin wireworms from soil smples olleted is time-onsuming (severl dys or more).
Pheromone-ited trps re muh esier nd simpler to use: the YF trp design proved to e exellent for severl lik eetle spp. in Europe nd North Ameri. However, the pheromone omposition should e identified!
Kép: Vuts J. The two speies we worked with: A. linetus A. proximus Determintion keys used to tell prt the speies: Rtio of width vs. length of thorx different & sutle differenes in genitl morphology Kép: Tóth M. : lous lterlis : lous medilis
GC nlysis of pheromone glnd extrt of A. linetus ws dominted y huge pek of gernyl otnote orresponding to literture dt 6 8 1 12 1 16 18 2 22 2 min gernyl otnote int. std.
Composition desried for West Ukrinin popultion ttrted no A. linetus. The ttrtion of A. ustultus is due to the frnesyl ompound, whih is its known pheromone. Totl ught Dereen, Hungry Apr 21 - Aug 5, 199 Gernyl otnote Neryl isovlerte E,E-frnesyl ette A. linetus A. ustultus + -- -- Desried s pheromone omponent y Borg-Krlson et l., 1988 (Swedish popultion), nd Siirde et l., 1993 (Est- Ukrinin nd Russin popultions) -- -- + -- + + Desried s pheromone omponents y Siirde et l., 1993 (West Ukrinin popultion). 3 195 93
Gernyl otnote showed only low to moderte field tivity on A. linetus. Totl ught Dereen, Hungry Apr 21 - Aug 5, 199 Gernyl otnote Neryl isovlerte E,E-frnesyl ette A. linetus A. ustultus + -- -- Desried s pheromone omponent y Borg-Krlson et l., 1988 (Swedish popultion), nd Siirde et l., 1993 (Est- Ukrinin nd Russin popultions) -- -- + -- + + Desried s pheromone omponents y Siirde et l., 1993 (West Ukrinin popultion). 3 195 93
Among omponents found in tre mounts in the glnd extrt, the ddition of gernyl utnote is highly synergisti in field trpping tests. Totl ught in test: 279 Rümlng, My 23 - August 1, 1997 Gernyl otnote Gernyl utnote E,E-frnesyl ette Gerniol Gernyl hexnote Neryl isovlerte A.linetus 1 3 2 1 1 1 - - - - 273 1-1 - - - 2 1 - - 1 - - 1 1 - - - 1-1 - - - - 1 1
This synergisti tivity of gernyl utnote ws lso onfirmed in further field tests. 1 men th + SE 8 6 2 Dereen, 1997 máj. 7. - júl 9. Totl ught: 22 Bit omposition (mg) 1 1 3 1,3 gernyl otnote gernyl utnote
The reltively high rtio of gernyl utnote in effiient its for A. linetus is surprising, s in pheromone glnd extrts this ompound ws found only in tres. 6 8 1 12 1 16 18 2 22 2 min gernyl otnote int. std. retention region of gernyl utyrte
Trps ited with A. linetus pheromone it (gernyl otnote + gernyl utnote) regulrly ught A. proximus in Portugl linetus thes proximus thes no th This ws surprising, euse sex pheromone omposition for A. proximus in literture: (E,E)- frnesyl ette + neryl isovlerte 99:1 [Ytsynin et l., Khim.Sel'sk.Khoz. Mosow, Khimiy, 33-35 (198)]
Agriotes proximus Agriotes linetus Men thes of A. proximus Men thes of A. linetus 1.9.8.7.6.5..3.2.1 Bit (mg) 1 2 3 5 6 7 8 9-3 3 3 3 3 3 3 3 - Brenits, Bulgri 25 Totl ught: 36 3.3 Hjdúöszörmény, Hungry 25 Totl ught: 36 12 1 8 6 2 gernyl utnote gernyl otnote neryl isovlerte (E,E)-frnesyl ette ette 2 6 8 1 12-3 3 3 3 Viro, Portugl 2 Totl ught: 1985 3 3 3 3 - - 3.3 Krsnodr, Russi 25 Totl ught: 61 A systemti field trpping test ws performed (2 sites for eh speies) ompring rtio rnge of the omponents identified y us nd the lend previously desried in literture for A. proximus.
Agriotes proximus Agriotes linetus Men thes of A. proximus Men thes of A. linetus 1.9.8.7.6.5..3.2.1 Bit (mg) 1 2 3 5 6 7 8 9-3 3 3 3 3 3 3 3 - Brenits, Bulgri 25 Totl ught: 36 3.3 Hjdúöszörmény, Hungry 25 Totl ught: 36 12 1 8 6 2 gernyl utnote gernyl otnote neryl isovlerte (E,E)-frnesyl ette ette 2 6 8 1 12-3 3 3 3 Viro, Portugl 2 Totl ught: 1985 3 3 3 3 - - 3.3 Krsnodr, Russi 25 Totl ught: 61 1) The lend previously desried in literture for A. proximus ws intive.
Agriotes sputtor Men thes of A. sputtor 5.5 3.5 3 2.5 2 1.5 1.5 Bit (mg) Men thes of A. ustultus 2 6 8 1 12 1 16 18 Krsnodr, Russi 25 Totl ught: 18-3 3 3 3 3 3 3 3 - Brenits, Bulgri 25 Totl ught: 3371 Agriotes ustultus 3.3.5 3.5 2.5 1.5.5 gernyl utnote gernyl otnote neryl isovlerte (E,E)-frnesyl ette 3 2 1 Hjdúöszörmény, Hungry 25 Totl ught: 11-3 3 3 3 Other spp. ptured: A. sputtor: gernyl utnote is min pheromone omponent A. ustultus: (E,E)-frnesyl ette is min pheromone omponent 3 3 3 3 - - 3.3
Agriotes proximus Agriotes linetus Men thes of A. proximus Men thes of A. linetus 1.9.8.7.6.5..3.2.1 Bit (mg) 1 2 3 5 6 7 8 9-3 3 3 3 3 3 3 3 - Brenits, Bulgri 25 Totl ught: 36 3.3 Hjdúöszörmény, Hungry 25 Totl ught: 36 12 1 8 6 2 gernyl utnote gernyl otnote neryl isovlerte (E,E)-frnesyl ette ette 2 6 8 1 12-3 3 3 3 Viro, Portugl 2 Totl ught: 1985 3 3 3 3 - - 3.3 Krsnodr, Russi 25 Totl ught: 61 2) For est ttrtion in oth speies oth gernyl utnote nd otnote ws neessry, in out equl mounts no differene etween the two speies.
Dose test: onfirmed the neessity of oth omponents for A. proximus Agriotes proximus 8 7 6 Koshv, Bulgri 25 Totl ught: 297 5 3 2 1 Bit (mg) 1 3 1 1 3 1 7 3-1 - 1 - - gernyl utnote gernyl otnote
Antennl responses for oth A. proximus nd A. linetus were reorded y n EAG pprtus ir flow with stimulus ntenn of A. proximus glss pillry eletrodes EAG-pek eliited y gernyl utnote
gernyl propionte gernyl utnote gerniol gernyl hexnote gernyl isovlerte gernyl otnote (E,E)-frnesyl ette (E,E)-frnesyil utyrte (E,E)-frnesol ir 15 1 5 stimulus men normlized response EAG response spetr of mle ntenne to numer of syntheti lik eetle pheromone omponents - no differene etween the two speies % 5 35 3 25 2 15 1 5 5 35 3 25 2 d e d f e g f Agriotes proximus Agriotes linetus
Antenne of oth speies responded etter to gernyl utnote % no differene etween the two speies men normlized responses 35 3 25 2 15 1 5 35 3 25 2 15 1 5 % ir (ontrol) ir (ontrol) Agriotes proximus P=,937 P=,2 P<,1 P<,1 P<,1 Agriotes linetus P=,9877 P=,386 P<,1 P=,121 P=,375 P<,1 P<,1 gernyl utnote gernyl otnote gernyl utnote gernyl otnote.1.1.1.1 1 1 dose (ug)
GC nlysis of voltile olletions from live ferl femle mv 1 A. linetus 5 X int. std. gernylotnote gernylutnote X mv A. proximus 5 25 X int. std. X X gernylotnote gernylutnote 2 6 8 1 12 1 16 18 2 22 2 min Speil thnks re due to Év Bálintné Csonk (PPI HAS, Budpest) for performing the olletions.
Preliminry moleulr omprisons on ferl speimens olleted in Bulgri (A. proximus) or Itly (A. linetus) [Speil thnks re due to Tmás Felföldi (ELTE, Budpest) for dvie nd diretions in performing moleulr studies] Gel eletrophoresis of PCR produts of ytohrome oxydse gene rrow shows 88 se pirlong DNA frtion Agriotes linetus Agriotes proximus negtive ontrol mrker
Preliminry moleulr omprisons on ferl speimens olleted in Bulgri (A. proximus) or Itly (A. linetus) [Speil thnks re due to Tmás Felföldi (ELTE, Budpest) for dvie nd diretions in performing moleulr studies] Comprison of DNA sequenes oding ytohrome oxidse: ferl insets olleted y us, or dt from literture dtse Kimur 2-prmeter model, Neighour-joining method Agriotes proximus (Bulgri) Agriotes linetus (Itly) Agriotes linetus DQ198566 Agriotes umintus DQ1561 Agriotes sputtor DQ15595.2 Melnotus riulosus EF275
Preliminry moleulr omprisons on ferl speimens olleted in Bulgri (A. proximus) or Itly (A. linetus) [Speil thnks re due to Tmás Felföldi (ELTE, Budpest) for dvie nd diretions in performing moleulr studies] Comprison of DNA sequenes oding ytohrome oxidse: ferl insets olleted y us, or dt from literture dtse A. proximus (Bulgri) A. linetus (Itly) A. linetus DQ198566 A. umintus DQ1561 A. sputtor DQ15595 M. riulosus EF275 A. proximus (Bulgri).2 A.linetus (Itly).5.3 A. linetus DQ198566.15.15.15.122.122.122.15 A. umint. A. sputtor DQ1561 DQ15595.156.156.158.199.173 Figures give rtio of differing vs. similr se pirs
Summry We found no signifint differene etween A. linetus nd A. proximus in: omposition of femle-produed pheromone (oth glnd extrts nd voltile olletions) eletrophysiologil responses of mles to syntheti pheromone omponents optiml field omposition of syntheti pheromone preliminrily in some moleulr spets Therefore txonomi revision of the two tx my eome worthwhile.